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53583 ^™

pKS3 [KS334] (ATCC^® 53583^™)

Organism: Escherichia coli (Migula) Castellani and Chalmers / Clone Type: Clone / Depositors: Harvard Univ., KL Strauch, Harvard Univ.

pKS3 [KS334] ATCC^® 53583^™

freeze-dried

For-Profit: $190.00 Non-Profit: $190.00

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Permits and Restrictions	View Permits
Designations	pKS3 [KS334]
Species	Escherichia coli (Migula) Castellani and Chalmers
Depositors	Harvard Univ., KL Strauch, Harvard Univ.
Applications	produces protein methyl-accepting chemotaxis protein I (inner membrane protein)
Vector	Construct size (kb): 6.0 DESCRIPTION OF VECTOR COMPONENT: Name of vector: pBR322 Intact vector size: 4.363 Type of vector: plasmid Cloning sites: EcoRI ClaI HindIII EcoRV BamHI SphI SalI XmaIII NruI BspMI BsmI StyI AvaI BalI BspMII PvuII Tth111I NdeI AflIII PpaI PstI PvuI ScaI SspI AatII Polylinker sites: Construction: pBR313 Host range: Escherichia coli Features (with orientation and position when available): marker(s): tetR replicon: pMB1 marker(s): ampR Cross references: DNA Seq. Acc.: J01749
Insert	DNA: genomic DESCRIPTION OF INSERT COMPONENT: Genome: Escherichia coli Gene symbol: phoA Gene name: alkaline phosphatase Contains complete coding sequence?: U Localization: 9 min Genome: Escherichia coli Gene symbol: tsr Gene name: methyl-accepting chemotaxis protein I (inner membrane protein) Contains complete coding sequence?: U Type of DNA: genomic Insert end: Insert end: Insert size (kb): Cross references: Gene product: methyl-accepting chemotaxis protein I (inner membrane protein) [tsr] Target Gene: methyl-accepting chemotaxis protein I (inner membrane protein)
Media	ATCC^® Medium 1896: 1065 plus ampicillin (100 mcg/ml) and XP (40 mcg/ml)
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Shipping Information	Distributed: freeze-dried
Comments	Restriction digests of the clone give the following sizes (kb): BamHI--uncut; EcoRI--4.4, 1.55, 0.35; PstI--6.0; HindIII--6.0. If the chimeric protein is degraded by proteases, the alkaline phosphatase becomes soluble in the periplasm and retains activity. The host chromosome contains an insertionally inactivated gene for the periplasmic protease htrA (degP), and thus shows reduced protease activity and increased stability of periplasmic proteins. pKS3 encodes a Tsr-alkaline phosphatase chimeric protein, with Tsr being N-terminal and anchoring the protein in the membrane. The alkaline phosphatase portion is exposed in the periplasm and retains activity.
Classification	Enterobacteriaceae, Escherichia
References	Beckwith JR, Strauch KL. Periplasmic protease mutants of Escherichia coli. US Patent 4,946,783 dated Aug 7 1990 Strauch KL, Beckwith J. An Escherichia coli mutation preventing degradation of abnormal periplasmic proteins. Proc. Natl. Acad. Sci. USA 85: 1576-1580, 1988. PubMed: 3278319
Disclosure	This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.

Bascic Documentation	Product Sheet Certificate of Analysis SDS
U.S. Patent	4,946,783
Disclosure	This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.